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Enviro-Chem Analytical, Inc. <br />685 West Gunnison Avenue, Suite 108 <br />Grand Junction, CO 81505 Outfall 001 <br />Sample Preuaration and Usage <br />2 <br />Sample information is contained in the table below. <br />Sample Collection <br />Feb. 26, 2008 11:00 am <br />Sam <br />Test Initiation and Renewal <br />Acute Toxicity Testing <br />West Elk Mine -Feb 2008 <br />2 Species <br />Sample Volume <br />6.0 gallons <br />Chain of Custody forms for the sample are contained in Appendix A. <br />Sample preparation followed test guidelines outlined by the USEPA. <br />Test Performance <br />Effluent concentrations of 100%, 50%, 25%, 13% and 6.5% were utilized during <br />the tests. Concentrations at less than 100% were obtained by diluting the <br />effluent samples with MHW made in the laboratory. All volume measurements <br />were made using graduated cylinders of minimum size. <br />Test temperature for both species was maintained at 20 +/- 1 C. Initial chemistry <br />measurements of pH, dissolved oxygen, temperature and conductivity were <br />performed for each effluent test concentration. Additionally, alkalinity and <br />hardness were performed for the MHW and 100% effluent sample. <br />Final chemistry measurements included temperature, pH, and dissolved oxygen <br />for each concentration and MHW control sample at the end of each renewal <br />period. <br />Fathead Minnows - <br />Four replicates per exposure concentration were tested with 10 organisms per <br />test chamber. Organisms were obtained from Aquatic Biosystems Inc. Fort <br />Collins, Colorado and were 3 days old at test initiation. Fish were impartially <br />distributed to each test chamber.Test chamber consisted of 500 mL disposable <br />plastic cups containing 250 mL of test solution. Fathead minnows were fed once <br />daily, 0.15 mL 2 hours prior to water renewal, newly hatched brine shrimp <br />(Artemis). Brine shrimp cysts were obtained from Aquatic Biosystems Inc, Fort <br />Collins, Colorado and were hatched in salt water overnight. Brine shrimp were <br />thoroughly rinsed prior to feeding to remove any residual salt. Test solution was <br />3 <br />